Untouched cd4 t cell isolation kit

Manufactured by Miltenyi Biotec

The Untouched CD4+ T cell isolation kit is a tool for the isolation of untouched CD4+ T cells from human peripheral blood mononuclear cells. The kit utilizes a magnetic bead-based negative selection approach to isolate the target cells without direct labeling.

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Lab products found in correlation

Facs vantage se diva cell sorter, by BD (2 mentions) Anti cxcr3 pe antibody, by R&D Systems (2 mentions) Cd4 cxcr3 cells, by Miltenyi Biotec (2 mentions) Anti pe microbeads, by Miltenyi Biotec (2 mentions) Anti pe beads, by Miltenyi Biotec (1 mentions) Rhtgf β1, by Thermo Fisher Scientific (1 mentions) Rmil 6, by R&D Systems (1 mentions) Rmil 2, by R&D Systems (1 mentions) Rmil 4, by R&D Systems (1 mentions) Anti ifn γ, by R&D Systems (1 mentions) Tepp 46, by Merck Group (1 mentions) Stattic, by Bio-Techne (1 mentions) Rapamycin, by Cayman Chemical (1 mentions) Rmil 23, by R&D Systems (1 mentions) Anti cd3ε cd28, by BD (1 mentions) Automacs magnetic cell sorter, by Miltenyi Biotec (1 mentions)

Spelling variants of this product

Isolation kits (10 citations) Cell isolation kits (5 citations) CD4 untouched kit (2 citations)

5 protocols using untouched cd4 t cell isolation kit

Helminth Regulation of Treg-mediated Cytokine Modulation

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To determine TGFβ cytokine generation of Treg-enriched vs Treg-depleted cultures, CD4+ T cells were purified from splenic and mesenteric lymph node (MLN) single cell suspensions of H. polygyrus-infected and uninfected Balb/C mice using CD4 T cell untouched isolation kit (Miltenyi Biotech) and separated into CD25 positive and CD25 negative fractions using anti-CD25 PE labeling, followed by magnetic separation with anti-PE beads (Miltenyi Biotech). Enrichment or depletion efficiency was >98% with these techniques (data not shown). To determine helminthic regulation of donor T cell IFNγ TNFα and interleukin 4 (IL4) cytokine output during GVHD, donor CD3+ T cells were sorted from total anti-CD3 FITC and anti-H2b PE stained splenocytes of uninfected and H. polygyrus-infected Balb/C recipients 6 days after GVHD induction using FACS Vantage SE DiVa cell sorter (Becton Dickinson).

Li Y., Chen H.L., Bannick N., Henry M., Holm A.N., Metwali A., Urban JF J.r., Rothman P.B., Weiner G.J., Blazar B.R., Elliott D.E, & Ince M.N. (2014). Intestinal Helminths Regulate Lethal Acute Graft Versus Host Disease and Preserve Graft Versus Tumor Effect in Mice. Journal of immunology (Baltimore, Md. : 1950), 194(3), 1011-1020.

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Helminth-Induced Regulation of T Cell Cytokines

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The expression of LAP and the production of TGFβ cytokine were assessed by purifying CD4⁺ T cells from splenic and mesenteric lymph nodes (MLN) of Hpb-infected and uninfected WT BALB/c, STAT6^–/–, STAT6 VT, furin^fl/fl, furin^fl/fl x CD4 Cre mice using the CD4 T Cell Untouched Isolation Kit (Miltenyi Biotech). This resulted in >98% enrichment for CD4+ T cells (data not shown). To assess helminthic regulation of cytokine production by donor T cells during GVHD, donor CD3⁺ T cells from uninfected and Hpb-infected WT BALB/c or STAT6^–/– BMT recipients were sorted from total splenocytes stained with anti-CD3 FITC and anti-H2^b PE 6 days after GVHD induction, using a FACS Vantage SE DiVa cell sorter (Becton Dickinson).

Li Y., Liu W., Guan X., Truscott J., Creemers J.W., Chen H.L., Pesu M., El Abiad R.G., Karacay B., Urban JF J.r., Elliott D.E., Kaplan M.H., Blazar B.R, & Ince M.N. (2018). STAT6 and Furin Are Successive Triggers for the Production of TGFβ by T Cells. Journal of immunology (Baltimore, Md. : 1950), 201(9), 2612-2623.

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T cell Differentiation and Modulation

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Naive CD4⁺CD25⁻ T cells were purified from LNs and spleen of WT C57BL/6, CD4^CrePkm2^fl/fl or control littermate (CD4^Cre and Pkm2^fl/fl) mice with the untouched CD4 T cell isolation kit (Miltenyi Biotec) and a biotinylated CD25 monoclonal antibody (eBioscience) by using an AutoMACS magnetic cell sorter (Miltenyi Biotec) according to the manufacturer’s protocol. Purified cells were activated with soluble anti-CD3ε:CD28 (both 1 µg/ml; BD Biosciences) on U-bottomed plates (10⁵/well). Skewing conditions were as follows: Th17, 2.5 ng/ml rhTGF-β1 (eBioscience) plus 20 ng/ml rmIL-6 (R&D Systems) with or without 20 ng/ml rmIL-23 (R&D Systems); Th1, rmIL-12, and rmIL-2 (both 20 ng/ml; R&D Systems); Th2, anti-IFN-γ (10 µg/ml), rmIL-4, and rmIL-2 (both 20 ng/ml; R&D Systems). For iT reg cell polarization, naive T cells were cultured with plate-bound CD3ε:CD28 (both 1 µg/ml; BD Biosciences) in the presence of 1 ng/ml rhTGF-β1 (eBioscience). When indicated, 0.1 µM rapamycin (Cayman Chemical), 100 µM TEPP-46 (Millipore), or 2 µM Stattic (Tocris) was used.

Damasceno L.E., Prado D.S., Veras F.P., Fonseca M.M., Toller-Kawahisa J.E., Rosa M.H., Públio G.A., Martins T.V., Ramalho F.S., Waisman A., Cunha F.Q., Cunha T.M, & Alves-Filho J.C. (2020). PKM2 promotes Th17 cell differentiation and autoimmune inflammation by fine-tuning STAT3 activation. The Journal of Experimental Medicine, 217(10), e20190613.

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Adoptive Transfer of Aged CD4+ T Cells

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Bulk CD4⁺ T cells were isolated from spleens and CLN from young and aged B6 mice using magnetic beads according to the manufacturer’s instructions (untouched CD4⁺ T cell isolation kit, Miltenyi Biotec, Auburn, CA). The CD4 enriched cell suspensions contained ≥ 90% CD4⁺ T cells as determined by flow cytometry (data not shown).
CD4⁺CXCR3⁺ cells were isolated from the spleens and CLN from young and aged B6 mice via a two-step procedure in which CD4⁺ T cells were pre-enriched by depletion of unwanted cells using the kit described above. The CD4⁺ T cell fraction was then incubated with the anti-CXCR3_PE antibody (R&D Systems, Minneapolis, MN) and then magnetically labeled with anti-PE microbeads and positively selected for CD4⁺CXCR3⁺ cells, according to the manufacturers’ instructions (Miltenyi Biotec, Auburn, CA). These cells were used to perform adoptive transfer experiments or collected for quantitative gene expression. Young and aged bulk CD4⁺T or CD4⁺CXCR3⁺ cells (2×10⁶) were transferred intraperitoneally (i.p.) into 6–10 week old T cell deficient RAG1KO mice. Experiments were performed five weeks after adoptive transfer of bulk CD4⁺T cells or CD4⁺CXCR3⁺ cells. Eyes were collected for goblet cell density analysis and LGs were collected for histology.

Bian F., Xiao Y., Barbosa F.L., de Souza R.G., Hernandez H., Yu Z., Pflugfelder S.C, & de Paiva C.S. (2019). Age-associated Antigen-Presenting Cell Alterations Promote Dry-Eye Inducing Th1 cells. Mucosal immunology, 12(4), 897-908.

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Adoptive Transfer of Aged CD4+ T Cells

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